| Type: | antibody |
| Identifier: | Alcam antibodyA |
| Entity Detected: | Alcam, activated leukocyte cell adhesion molecule ( MGI:1313266) |
| Antigen: | sense strand is shown
>Alcam antibodyA
MASKVSPSCRLVFCLLISAAVLRPGLGWYTVNSAYGDTIVMPCRLDVPQNLMFGKWKYEKPDGSPVFIAF
RSSTKKSVQYDDVPEYKDRLSLSENYTLSIANAKISDEKRFVCMLVTEDNVFEAPTLVKVFKQPSKPEIV
NKAPFLETDQLKKLGDCISRDSYPDGNITWYRNGKVLQPVEGEVAILFKKEIDPGTQLYTVTSSLEYKTT
RSDIQMPFTCSVTYYGPSGQKTIYSEQEIFDIYYPTEQVTIQVLPPKNAIKEGDNITLQCLGNGNPPPEE
FMFYLPGQPEGIRSSNTYTLTDVRRNATGDYKCSLIDKRNMAASTTITVHYLDLSLNPSGEVTKQIGDTL
PVSCTISASRNATVVWMKDNIRLRSSPSFSSLHYQDAGNYVCETALQEVEGLKKRESLTLIVEGKPQIKM
TKKTDPSGLSKTIICHVEGFPKPAIHWTITGSGSVINQTEESPYINGRYYSKIIISPEENVTLTCTAENQ
LERTVNSLNVSAISIPEHDEADDISDENREKVNDQAKLIVGIVVGLLLAALVAGVVYWLYMKKSKTASKH
VNKDLGNMEENKKLEENNHKTEA
|
| | aa 1 - aa 583 of NP_033785.1 |
| Notes: | The anti-ALCAM antibody used in this study by Hirata et al., 2006 [PMID:17409743] is described as "rat anti-mouse ALCAM (ALC-48; 1:1,000 in use)". Generation of mAbs against murine ALCAM is described by the authors as follows: "HEK293T and BaF3 cells were transfected with MIE-ALCAM, using FuGENE 6 Transfection Reagent (Roche Diagnostics Japan) following the manufacturer's instructions. 293T cells positive for eGFP expression were sorted by FACS Aria (BD Biosciences, San Jose, Calif., USA) and immunized into Wistar rats. The spleen cells from the immunized rats were harvested and fused with the murine myeloma cell line P3-X63-Ag8-U1, as previously described (Sudo et al., 1993 [PMID:8415665] ). For screening the hybridoma clone producing specific antibodies against ALCAM, supernatant from hybridoma cell cultures was reacted with exogenous ALCAM antigen expressed on BaF3 cells by stable transfection with Mie- ALCAM (BaF3-mie-ALCAM), followed by detection with goat anti-rat IgG antibodies conjugated with phycoerythrin (Beckman Coulter, Fullerton, Calif., USA) by FACS Aria and analyzed with FACS Diva (BD Biosciences) and FlowJo (Tree Star, Ashland, Oreg., USA) software."
MIE-ALCAM was made as follows: "The entire coding sequence of murine ALCAM (NM_009655, nucleotides 1609-3360) was amplified as an EcoRI-BamHI fragment by reverse transcription-polymerase chain reaction from total RNA extracts of embryonic day (E) 12.5 fetus livers. The fragments were cloned into Mie ((MSCV, murine stem cell virus)-IRES (internal ribosome entry site)-enhanced green fluorescent protein (eGFP)) retroviral vector (Kawamata et al., 2002 [PMID:12032823] ) at the EcoRI-BamHI sites".
Editors Note: NM_009655.1, nucleotides 1609-3360 covers the entire ORF from the first to last amino acid. |
| Antibody Type: | monoclonal |
| Raised In: | rat |
icon to keep this page displayed.)
